Production of infectious hepatitis C virus of various genotypes in cell cultures

Takanobu Kato, Takuya Matsumura, Theo Heller, Satoru Saito, Ronda K. Sapp, Krishna Murthy, Takaji Wakita, T. Jake Liang

Research output: Contribution to journalArticle

Abstract

A unique hepatitis C virus (HCV) strain JFH-1 has been shown to replicate efficiently in cell culture with production of infectious HCV. We previously developed a DNA expression system containing HCV cDNA flanked by two self-cleaving ribozymes to generate HCV particles in cell culture. In this study, we produced HCV particles of various genotypes, including 1a (H77), 1b (CG1b), and 2a (J6 and JFH-1), in the HCV-ribozyme system. The constructs also contain the secreted alkaline phosphatase gene to control for transfection efficiency and the effects of culture conditions. After transfection into the Huh7-derived cell line Huh7.5.1, continuous HCV replication and secretion were confirmed by the detection of HCV RNA and core antigen in the culture medium. HCV replication levels of strains H77, CG1b, and J6 were comparable, whereas the JFH-1 strain replicates at a substantially higher level than the other strains. To evaluate the infectivity in vitro, the culture medium of JFH-1-transfected cells was inoculated into naive Huh7.5.1 cells. HCV proteins were detected by immunofluorescence 3 days after inoculation. To evaluate the infectivity in vivo, the culture medium from HCV genotype 1b-transfected cells was inoculated into a chimpanzee and caused a typical course of HCV infection. The HCV 1b propagated in vitro and in vivo had sequences identical to those of the HCV genomic cDNA used for cell culture transfection. The development of culture systems for production of various HCV genotypes provides a valuable tool not only to study the replication and pathogenesis of HCV but also to screen for antivirals.

Original languageEnglish (US)
Pages (from-to)4405-4411
Number of pages7
JournalJournal of Virology
Volume81
Issue number9
DOIs
StatePublished - May 2007
Externally publishedYes

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Hepatovirus
hepatitis A
Hepatitis C virus
Hepacivirus
cell culture
Cell Culture Techniques
Genotype
genotype
transfection
Transfection
Culture Media
Catalytic RNA
culture media
Virus Replication
virus replication
virion
Virion
pathogenicity
Complementary DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Kato, T., Matsumura, T., Heller, T., Saito, S., Sapp, R. K., Murthy, K., ... Liang, T. J. (2007). Production of infectious hepatitis C virus of various genotypes in cell cultures. Journal of Virology, 81(9), 4405-4411. https://doi.org/10.1128/JVI.02334-06

Production of infectious hepatitis C virus of various genotypes in cell cultures. / Kato, Takanobu; Matsumura, Takuya; Heller, Theo; Saito, Satoru; Sapp, Ronda K.; Murthy, Krishna; Wakita, Takaji; Liang, T. Jake.

In: Journal of Virology, Vol. 81, No. 9, 05.2007, p. 4405-4411.

Research output: Contribution to journalArticle

Kato, T, Matsumura, T, Heller, T, Saito, S, Sapp, RK, Murthy, K, Wakita, T & Liang, TJ 2007, 'Production of infectious hepatitis C virus of various genotypes in cell cultures', Journal of Virology, vol. 81, no. 9, pp. 4405-4411. https://doi.org/10.1128/JVI.02334-06
Kato T, Matsumura T, Heller T, Saito S, Sapp RK, Murthy K et al. Production of infectious hepatitis C virus of various genotypes in cell cultures. Journal of Virology. 2007 May;81(9):4405-4411. https://doi.org/10.1128/JVI.02334-06
Kato, Takanobu ; Matsumura, Takuya ; Heller, Theo ; Saito, Satoru ; Sapp, Ronda K. ; Murthy, Krishna ; Wakita, Takaji ; Liang, T. Jake. / Production of infectious hepatitis C virus of various genotypes in cell cultures. In: Journal of Virology. 2007 ; Vol. 81, No. 9. pp. 4405-4411.
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